LOCUS       sequence-319647-        8478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses KCNK7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8478)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8478)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: IDG_KCNK7_OE_1
FEATURES             Location/Qualifiers
     source          1..8478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    531..555
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1312..1336)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1338..1379
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1421..2009
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1474..1494)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1892..1903)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2081..2314
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2392..2513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2429..2448)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2483..2502
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2553..2688
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2708..2727)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2709..2727)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2737..2754)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2737..2753)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2746..2768)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2895..3350
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2982..3001)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3192..3213
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3376..3480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3481..3549,3550..4341)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3699..3718)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4512..5100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5001..5020
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5254..5271
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5388..5409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5424..5429,5430..5447,5448..5454)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5462..5478
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5467..5489
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5486..5502
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5486..5502
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5521..5541
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5523..5541
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5569..5795
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5796..5976
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6023..6148
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6641..6874
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7059..7103
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        7278..7778
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7697..7715
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7790..8185
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8245..8362
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact gtcgggatca acaagtttgt
      541 acaaaaaagc aggcaccatg gggggtctaa ggccctggtc ccgatacggg ctcctggttg
      601 tggcccactt gctggccctg gggcttgggg ctgtggtgtt ccaggccctg gaggggcctc
      661 ctgcatgcag gcttcaggct gagctcaggg cagagctggc agccttccag gcagagcata
      721 gggcctgcct gccacccgga gctctggaag agctgctggg cactgccctg gccacccagg
      781 cccatggggt ctccaccctg ggcaacagct cagagggcag gacctgggac cttccctcag
      841 ccctgctctt cgctgccagc atcctcacca ccacaggtta tggccacatg gccccactat
      901 cgccaggcgg aaaggccttc tgcatggtct atgcagccct ggggctgcca gcctccttag
      961 ctctcgtggc caccctgcgc cattgcctgc tgcctgtgct cagccgccca cgtgcctggg
     1021 tagcggtcca ctggcagctg tcaccggcca gggctgcgct gctgcaggca gttgcactgg
     1081 gactgctggt ggccagcagc tttgtgctgc tgccagcgct ggtgctgtgg ggccttcagg
     1141 gcgactgcag cctgctgggg gccgtctact tctgcttcag ctcgctcagc accattggcc
     1201 tggaggactt gctgcccggc cgcggccgca gcctgcaccc cgtgatttac cacctgggcc
     1261 agctcgcact tcttggtgga gggacctcac tccagggcac ggcgtgggag gggacagctt
     1321 tcttgtacaa agtggttggt aagcctatcc ctaaccctct cctcggtctc gattctacgt
     1381 agtaatgagc tagcgctaac cggtggcgcg ttaagtcgac aatcaacctc tggattacaa
     1441 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     1501 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     1561 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     1621 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     1681 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     1741 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     1801 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     1861 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     1921 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     1981 tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca
     2041 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     2101 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     2161 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     2221 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     2281 agaccctttt agtcagtgtg gaaaatctct agcagtacgt atagtagttc atgtcatctt
     2341 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     2401 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2461 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     2521 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2581 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2641 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
     2701 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     2761 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     2821 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     2881 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2941 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3001 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3061 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3121 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3181 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3241 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3301 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     3361 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3421 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3481 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3541 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3601 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3661 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3721 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3781 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3841 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3901 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3961 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4021 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4081 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4141 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4201 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4261 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4321 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4381 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4441 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4501 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4561 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4621 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4681 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4741 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4801 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4861 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4921 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4981 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5041 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5101 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5161 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5221 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5281 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5341 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     5401 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     5461 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     5521 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
     5581 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
     5641 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
     5701 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
     5761 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
     5821 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5881 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5941 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     6001 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     6061 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     6121 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     6181 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     6241 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     6301 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     6361 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     6421 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     6481 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     6541 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     6601 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     6661 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     6721 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     6781 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     6841 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     6901 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     6961 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     7021 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     7081 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     7141 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     7201 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     7261 caaccccgag gggacccttg cgccttttcc aaggcagccc tgggtttgcg cagggacgcg
     7321 gctgctctgg gcgtggttcc gggaaacgca gcggcgccga ccctgggtct cgcacattct
     7381 tcacgtccgt tcgcagcgtc acccggatct tcgccgctac ccttgtgggc cccccggcga
     7441 cgcttcctgc tccgccccta agtcgggaag gttccttgcg gttcgcggcg tgccggacgt
     7501 gacaaacgga agccgcacgt ctcactagta ccctcgcaga cggacagcgc cagggagcaa
     7561 tggcagcgcg ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg
     7621 agagcagcgg ccgggaaggg gcggtgcggg aggcggggtg tggggcggta gtgtgggccc
     7681 tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg
     7741 gctccctcgt tgaccgaatc accgacctct ctccccaggg ggtaccacca tggccaagcc
     7801 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     7861 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     7921 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     7981 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     8041 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     8101 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     8161 gccctctggt tatgtgtggg agggcctgca gctgcagtag taagaattct agatcttgag
     8221 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
     8281 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     8341 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg
     8401 ccggctcgag ggggcccggg gttattaata gtaatcaatt acggggtcat tagttcatag
     8461 cccatatatg gagttccg
//
